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Literature summary for 3.2.1.31 extracted from
- Rapid evolution of beta-glucuronidase specificity by saturation mutagenesis of an active site loop (2004), J. Biol. Chem., 279, 26462-26468.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene gusA, expression of His-tagged wild-type and mutant enzymes in Escherichia coli | Escherichia coli |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D531E/S557V/N566S/G601S | saturation mutagenesis, mutant 1.13, altered substrate specificity compared to the wild-type enzyme | Escherichia coli |
| additional information | rapid evolution of beta-glucuronidase specificity by saturation mutagenesis of an active site loop, DNA shuffling of point mutations, construction of diverse mutants with mutation of residues 557, 566, and 568, the mutants show increased activity with beta-D-xylopyranoside and reduced activity with beta-D-glucuronide, overview | Escherichia coli |
| S22N/G81S/K257E/T509A/S557P/N566S/K568Q/Q598R/stop604W | saturation mutagenesis, mutant 1.15, altered substrate specificity compared to the wild-type enzyme | Escherichia coli |
| S557I/N566A/K568R/A580V | saturation mutagenesis, mutant 1.16, altered substrate specificity compared to the wild-type enzyme | Escherichia coli |
| S557Q/N566K/K568S/Q598stop | saturation mutagenesis, mutant 1.2, altered substrate specificity compared to the wild-type enzyme | Escherichia coli |
| V473A/S557P/N566S/K568Q | saturation mutagenesis, mutant 4.7, altered substrate specificity compared to the wild-type enzyme | Escherichia coli |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | kinetics of mutant enzymes, overview | Escherichia coli | |
| 0.2 | - |
4-nitrophenyl beta-D-xylopyranoside | wild-type enzyme | Escherichia coli |  |
| 0.36 | - |
4-nitrophenyl beta-D-glucuronide | wild-type enzyme | Escherichia coli | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Escherichia coli | - |
gene gusA | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged wild-type and mutant enzymes from Escherichia coli by nickel affinity chromatography to homogeneity | Escherichia coli |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 4-nitrophenyl beta-D-glucuronide + H2O | - |
Escherichia coli | 4-nitrophenol + beta-D-glucuronic acid | - |
? | |
| 4-nitrophenyl beta-D-xylopyranoside + H2O | preferred substrate | Escherichia coli | 4-nitrophenol + beta-D-xylopyranose | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| GUS | - |
Escherichia coli |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.001 | - |
4-nitrophenyl beta-D-glucuronide | wild-type enzyme | Escherichia coli | |
| 68 | - |
4-nitrophenyl beta-D-xylopyranoside | wild-type enzyme | Escherichia coli | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.6 | - |
assay at | Escherichia coli |
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